The present invention relates to a novel family of purified proteins designated BMP-3 proteins and processes for obtaining them. These proteins may be used to induce bone and/or cartilage formation and in wound healing and tissue repair.
BMP-3 proteins are produced by cloturing a cell transformed with a cDNA substantially as shown in Table II and recovering from the culture medium a protein containing substantially the 96 amino acid sequence as shown in Table II from amino acid # 377 through amino acid # 472.
Some members of the BMP-3 protein family are further characterized by the ability of 200 nanograms of the BMP-3 protein to score at least +2 in the Rosen-modified Sampath-Reddi assay of bond and/or cartilage formation described in Example III.
Another aspect of the invention provides pharmaceutical compositions containing a therapeutically effective amount of a BMP-3 protein in admixture with a pharmaceutically acceptable vehicle or carrier. The compositions may be used for bone and/or cartilage formation BMP-3 compositions may also be used for wound healing and tissue repair. Compositions of the invention may further include other therapeutically useful agents such as the BMP proteins BMP-1, BMP-2A, and BMP-2B disclosed respectively in coowned and concurrently filed pending U.S. patent applications Ser. No. 179,101 and Ser. No. 179,100. Other therapeutically useful agents include growth factors such as epidermal growth factor (EGF), fibroblast growth factor (FGF), and transforming growth factor (TGF). The compositions may also include an appropriate matrix, for instance, for supporting the compositions and providing a surface for bone and/or cartilage growth. The compositions may be employed in methods for treating a number of bone defects and periodontal disease and various types of wounds. These methods, according to the invention, entail administering to a patient needing such bone and/or cartilage formation, wound healing, or tissue repair an effective amount of a novel BMP-3 protein of the present invention. These methods may also entail the administration of a BMP-3 protein of the invention in conjunction with at least one of the novel BMP proteins disclosed in co-owned applications described above. In addition, these methods may also include administration of BMP-3 with other growth factors.
Still a further aspect of the invention are DNA sequences coding on expression for a BMP-3 protein. Such sequences include the sequence of nucleotides in a 5xe2x80x2 to 3xe2x80x2 direction illustrated in Tables I A and I B and II or DNA sequences which hybridize under stringent conditions with the DNA sequences of Tables I A and I B and II and encode a protein having the ability of 200 nanograms of the protein to score at least +2 in the Rosen-modified Sampath-Reddi assay of bone and/or cartilage formation described in Example III. Finally, allelic or other variations of the sequences of Tables I A and I B and II, whether such nucleotide changes result in changes in the peptide sequence or not, are also included in the present invention.
Still a further aspect of the invention is a vector containing a DNA sequence as described above in operative association with an expression control sequence therefor. Such vector may be employed in a novel process for producing a BMP-3 protein of the invention in which a cell line transformed with a DNA sequence encoding expression of a BMP-3 protein in operative association with an expression control sequence therefor, is cultured in a suitable culture medium and a BMP-3 protein is isolated and purified therefrom. This claimed process may employ a number of known cells both prokaryotic and eukaryotic as host cells for expression of the polypeptide.
Other aspects and advantages of the present invention will be apparent upon consideration of the following detailed description and preferred embodiments thereof.